MSPD-HPLC法分析云南红豆杉中的10-DAB Ⅲ和紫杉醇

Determination of 10-DAB Ⅲ and Taxol in Taxus yunnanensis by MSPD-HPLC

关键词:  云南红豆杉, 10-去乙酰基巴卡亭 Ⅲ, 紫杉醇, 基质固相萃取, 高效液相色谱分析

Key words:   Taxus yunnanensis, 10-DAB Ⅲ, Taxol, MSPD, HPLC

摘要:

为建立云南红豆杉(Taxus yunnanensis)中10-去乙酰基巴卡亭Ⅲ(10-DAB Ⅲ)和紫杉醇(Taxol)的分析方法,该研究采用基质固相萃取-高效液相色谱分析(MSPD-HPLC)对云南红豆杉中的10-DAB Ⅲ及Taxol进行定量分析,探究硅胶、佛罗里硅土、酸性氧化铝、中性氧化铝、碱性氧化铝、C₁₈、C₁₈-ME、C₁₈-G1、C₁₈-HC、Diol、Xamide、Xion 共12种样品预处理分散剂及各种分散剂的质量、洗脱剂的种类、洗脱剂的浓度、洗脱剂体积对两种成分分析的影响,并对优化后的条件进行方法学验证,同时与超声提取、热回流提取预处理方法进行比较分析证明其有效性。结果表明:(1)12种固相分散剂中,碱性氧化铝具有良好的目标化合物萃取物检出量,并且当碱性氧化铝与样品的质量比为3:1、6 mL甲醇为洗脱剂洗脱时,10-DAB Ⅲ和Taxol的萃取检出量较高。(2)建立的云南红豆杉枝叶中10-DAB Ⅲ和Taxol的分析方法具有良好的线性关系(r≥0.999 9),10-DAB Ⅲ和Taxol的检出限和定量限分别介于0.023 9～0.030 1 μg·mL^-1和0.142～0.178 μg·mL^-1之间; 各目标分析物平均加样回收率在93.6%～109.0%之间。(3)比较分析显示,3种方法对2种紫杉烷类化合物的萃取检出量基本无差别,但MSPD法溶剂消耗少、操作简单,分析时间短,净化效率高,可应用于云南红豆杉原料的快速分析。该研究以碱性氧化铝为分散剂应用于云南红豆杉紫杉烷类化合物的萃取,建立了快速、高效的云南红豆杉中10-DAB Ⅲ和紫杉醇的分析方法,为云南红豆杉中的紫杉烷类化合物的定量分析提供了依据。

Abstract:

This research aimed to establish an analytical method for the quantification of 10-deacetylbaccatin Ⅲ(10-DAB Ⅲ)and Taxol in Taxus yunnanensis. The method of matrix solid phase dispersion coupled with high performance liquid chromatography(MSPD-HPLC)was used for the respective quantification of 10-DAB Ⅲ and Taxol of T. yunnanensis. The influences of various parameters were investigated, including 12 types of solid-phase dispersants(silica gel, florisil, acid alumina, neutral alumina, alkaline alumina, C₁₈, C₁₈-ME, C₁₈-G1, C₁₈-HC, Diol, Xamide, Xion), the mass of the dispersants, and the type, concentration, and volume of the eluent on the analysis of the two components. After optimization of these conditions, the methodology was validated. Additionally, a comparative analysis was conducted with the conventional ultrasonic extraction and hot reflux extraction pretreatment methods to ascertain their effectiveness of our novel method. The results were as follows:(1)Among the 12 solid-phase dispersants examined, alkaline alumina yielded a superior extraction detection rate for 10-DAB Ⅲ and Taxol when used in a 3:1 ratio with the sample mass and 6 mL of methanol as the eluent.(2)The developed 10-DAB Ⅲ and Taxol analysis method in T. yunnanensis demonstrated that excellent linearity(r≥0.999 9)with limit of detection(LOD)and limit of quantification(LOQ)for 10-DAB Ⅲ and Taxol ranging from 0.023 9 to 0.030 1 μg·mL^-1 and from 0.142 to 0.178 μg·mL^-1, respectively. The average recoveries of the target analytes varied between 93.6% and 109.0%.(3)A comparative analysis revealed that negligible differences in extraction detection of the two taxanes between the three methods. However, the MSPD method stood out due to its lower solvent consumption, simple operation, short analysis time and high purification efficiency, making it more suitable for the rapid analysis of T. yunnanensis raw materials. Therefore, this study presents a rapid and efficient analytical method for the extraction and analysis of 10-DAB Ⅲ and Taxol in T. yunnanensis, using alkaline alumina as a dispersant, providing a significant contribution to the quantitative analysis of taxanes in this species.